Title
Biomarkers in cervical screening: quantitative reverse transcriptase PCR analysis of P16INK4a expressionBiomarkers in cervical screening: quantitative reverse transcriptase PCR analysis of P16INK4a expression
Author
Faculty/Department
Faculty of Medicine and Health Sciences
Research group
Laboratory of cell biology and histology
Laboratory for Microbiology, Parasitology and Hygiene (LMPH)
Faculty of Medicine and Health Sciences - other
Publication type
article
Publication
Oxford,
Subject
Human medicine
Source (journal)
European journal of cancer prevention. - Oxford
Volume/pages
19(2010):1, p. 35-41
ISSN
0959-8278
ISI
000272952200006
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Molecular insights into the human papillomavirus (HPV)-induced cervical carcinogenesis led to the discovery of biomarkers for cervical disease. The detection of cellular proteins that are overexpressed by HPV-infected cells, such as tumor suppressor protein p16INK4a, might play an important role in future cervical cancer screening strategies. P16INK4a immunostaining correlates with the severity of cytological and histological abnormalities, but shows some methodological shortcomings such as the lack of standardized methodology and interobserver variability. This study evaluated quantitative reverse transcriptase PCR (RT-PCR) as an alternative tool to analyze p16INK4a overexpression as a biomarker for transforming HPV-infections in a liquid-based cervical cytology (LBC) setting. Sixty LBC samples, divided in three groups based on their cytological diagnosis, were subjected to HPV typing and analysis of p16INK4a expression by immunocytochemistry and RT-PCR. The analytical sensitivity of the RT-PCR was determined by spiking HeLa and HaCaT cells. P16INK4a expression measured by RT-PCR did not correlate with the cytological diagnosis or HPV status (HPV-positivity, infection type and HPV16-positivity). The spiking experiment proved that, to detect increased biomarker expression by RT-PCR, about 1.0% dysplastic cells is required within a pool of normal keratinocytes. In conclusion, RT-PCR analysis of biomarker expression is not appropriate for cervical screening purposes. In typical LBC samples, the biomarker transcripts of the dysplastic cells are diluted by the RNA of the normal cells in such a manner that their overexpression cannot be detected by RT-PCR.
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